pMSCV16URP-U6-sh-HTS6-UbiC-TagRFP-2A-Puro

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HomePlasmidsViral Expression & Packaging VectorspMSCV16URP-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
SgrAI (7424) SspI (6685) XmnI (6480) ScaI (6361) AflIII - PciI (4988) HindIII (4757) AvrII (4741) SfiI (4694) HincII (3669) BbvCI (209) AscI (369) SpeI (661) BglII (1410) BbsI - BpiI (1683) BbsI - BpiI (1709) PaeR7I - PspXI - XhoI (1732) BsgI (1777) EcoRI (1849) AgeI (1858) BstBI (1867) AleI (2284) PflMI (2889) PshAI (2923) BamHI (2995) BsiWI (3117) BspEI * (3174) RsrII (3177) SalI (3667) pMSCV16URP-U6-sh-HTS6- UbiC-TagRFP-2A-Puro 7602 bp
SgrAI  (7424)
1 site		 C R C C G G Y G G Y G G C C R C
Efficient cleavage requires at least two copies of the SgrAI recognition sequence.
SspI  (6685)
1 site		 A A T A T T T T A T A A
XmnI  (6480)
1 site		 G A A N N N N T T C C T T N N N N A A G
ScaI  (6361)
1 site		 A G T A C T T C A T G A
AflIII  (4988)
1 site		 A C R Y G T T G Y R C A
Sticky ends from different AflIII sites may not be compatible.
PciI  (4988)
1 site		 A C A T G T T G T A C A
PciI is inhibited by nonionic detergents.
HindIII  (4757)
1 site		 A A G C T T T T C G A A
AvrII  (4741)
1 site		 C C T A G G G G A T C C
SfiI  (4694)
1 site		 G G C C N N N N N G G C C C C G G N N N N N C C G G
Efficient cleavage requires at least two copies of the SfiI recognition sequence.
Sticky ends from different SfiI sites may not be compatible.
HincII  (3669)
1 site		 G T Y R A C C A R Y T G
BbvCI  (209)
1 site		 C C T C A G C G G A G T C G
AscI  (369)
1 site		 G G C G C G C C C C G C G C G G
SpeI  (661)
1 site		 A C T A G T T G A T C A
BglII  (1410)
1 site		 A G A T C T T C T A G A
BbsI  (1683)
2 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BpiI  (1683)
2 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BpiI sites may not be compatible.
BbsI  (1709)
2 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
BpiI  (1709)
2 sites		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BpiI sites may not be compatible.
PaeR7I  (1732)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
PspXI  (1732)
1 site		 V C T C G A G B B G A G C T C V
XhoI  (1732)
1 site		 C T C G A G G A G C T C
BsgI  (1777)
1 site		 G T G C A G ( N ) 14 N N C A C G T C ( N ) 14
Efficient cleavage requires at least two copies of the BsgI recognition sequence.
Sticky ends from different BsgI sites may not be compatible.
For full activity, add fresh S-adenosylmethionine (SAM).
EcoRI  (1849)
1 site		 G A A T T C C T T A A G
AgeI  (1858)
1 site		 A C C G G T T G G C C A
BstBI  (1867)
1 site		 T T C G A A A A G C T T
AleI  (2284)
1 site		 C A C N N N N G T G G T G N N N N C A C
PflMI  (2889)
1 site		 C C A N N N N N T G G G G T N N N N N A C C
Sticky ends from different PflMI sites may not be compatible.
PshAI  (2923)
1 site		 G A C N N N N G T C C T G N N N N C A G
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
BamHI  (2995)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
BsiWI  (3117)
1 site		 C G T A C G G C A T G C
BsiWI is typically used at 55°C, but is 50% active at 37°C.
BspEI  (3174)
1 site		 T C C G G A A G G C C T
* Blocked by Dam methylation.
RsrII  (3177)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
SalI  (3667)
1 site		 G T C G A C C A G C T G
TagRFP
2284 .. 2994  =  711 bp
237 amino acids  =  26.7 kDa
Product: monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)
mammalian codon-optimized
TagRFP
2284 .. 2994  =  711 bp
237 amino acids  =  26.7 kDa
Product: monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)
mammalian codon-optimized
T2A
3001 .. 3054  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
T2A
3001 .. 3054  =  54 bp
18 amino acids  =  1.8 kDa
Product: 2A peptide from Thosea asigna virus capsid protein
Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides.
PuroR
3061 .. 3660  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
PuroR
3061 .. 3660  =  600 bp
199 amino acids  =  21.5 kDa
Product: puromycin N-acetyltransferase
confers resistance to puromycin
AmpR
5808 .. 6668  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5808 .. 6599  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5808 .. 6668  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   6600 .. 6668  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
5808 .. 6668  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
5049 .. 5637  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5049 .. 5637  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
5' LTR
7601 .. 515  =  517 bp
5' long terminal repeat from murine embryonic stem cell virus
5' LTR
7601 .. 515  =  517 bp
5' long terminal repeat from murine embryonic stem cell virus
3' LTR (ΔU3)
3736 .. 4161  =  426 bp
self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus
3' LTR (ΔU3)
3736 .. 4161  =  426 bp
self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus
gag (truncated)
987 .. 1403  =  417 bp
truncated MMLV gag gene lacking the start codon
gag (truncated)
987 .. 1403  =  417 bp
truncated MMLV gag gene lacking the start codon
UbiC promoter
1872 .. 2271  =  400 bp
human ubiquitin C promoter
UbiC promoter
1872 .. 2271  =  400 bp
human ubiquitin C promoter
MESV Ψ
579 .. 920  =  342 bp
packaging signal of murine embryonic stem cell virus
MESV Ψ
579 .. 920  =  342 bp
packaging signal of murine embryonic stem cell virus
SV40 promoter
4427 .. 4756  =  330 bp
SV40 enhancer and early promoter
SV40 promoter
4427 .. 4756  =  330 bp
SV40 enhancer and early promoter
U6 promoter
1438 .. 1686  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
U6 promoter
1438 .. 1686  =  249 bp
RNA polymerase III promoter for human U6 snRNA (Domitrovich & Kunkel, 2003)
AmpR promoter
6669 .. 6773  =  105 bp
AmpR promoter
6669 .. 6773  =  105 bp
SV40 ori
4607 .. 4742  =  136 bp
SV40 origin of replication
SV40 ori
4607 .. 4742  =  136 bp
SV40 origin of replication
cloning site
1684 .. 1713  =  30 bp
shRNA cloning site
cloning site
1684 .. 1713  =  30 bp
shRNA cloning site
ORF:  2284 .. 3660  =  1377 bp
ORF:  458 amino acids  =  50.3 kDa
ORF:  5938 .. 6204  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  7054 .. 7350  =  297 bp
ORF:  98 amino acids  =  11.7 kDa
ORF:  3985 .. 4275  =  291 bp
ORF:  96 amino acids  =  10.5 kDa
ORF:  2070 .. 2339  =  270 bp
ORF:  89 amino acids  =  10.0 kDa
ORF:  2397 .. 2642  =  246 bp
ORF:  81 amino acids  =  9.1 kDa
ORF:  5808 .. 6668  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  362 .. 637  =  276 bp
ORF:  91 amino acids  =  9.9 kDa
ORF:  674 .. 934  =  261 bp
ORF:  86 amino acids  =  9.5 kDa
ORF:  7118 .. 7348  =  231 bp
ORF:  76 amino acids  =  7.5 kDa
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Individual Sequences & Maps

pAAV-AC-GFP
pLenti6.2 N-Lumio V5-DEST
pLVX-rHom-1
pRetroX-Tet3G
pAAV-AC-Myc-DDK
pLenti6.2 V5-DEST
pLVX-rHom-Nuc1
pRetroX-TetOne
pAAV-AC-RFP
pLenti6.3 TO V5-DEST
pLVX-rHom-Sec1
pRetroX-TetOne-Puro
pAAV2-EF1a-tGFP-WPRE
pLenti6.3 V5-DEST
pLVX-Tet-Off Advanced
pRetroX-Tight-Hyg
pAd BLOCK-iT-DEST
pLenti6.4 R4R2 V5-DEST
pLVX-Tet-On Advanced
pRetroX-Tight-Pur
pAd CMV V5-DEST
pLenti6 BLOCK-iT-DEST
pLVX-Tet3G
pRetroX-TRE3G
pAd PL-DEST
pLenti6 capTEV-CT-DEST
pLVX-TetOne
pRFP-C-RS
pAdenoX-CMV Linear
pLenti6 capTEV-NT-DEST1
pLVX-TetOne-Puro
pRFP-CB-shLenti
pAdenoX-DsRed-Express Linear
pLenti6 TR
pLVX-Tight-Puro
pRS
pAdenoX-PRLS Linear
pLenti6 UbC V5-DEST
pLVX-TRE3G
pRSGC1-U6-sg-HTS6-CMV-Cas9-2A-Puro
pAdenoX-PRLS-DsRed-Express Linear
pLenti6 V5-DEST
pLVX-TRE3G-Hom1
pRSGC2-U6-sg-HTS6-UbiC-Cas9-2A-Puro
pAdenoX-PRLS-ZsGreen1 Linear
pLenti7.3 V5-DEST
pLVX-TRE3G-IRES
pRSI12-U6-sh-HTS4-UbiC-TagRFP-2A-Puro
pAdenoX-Tet3G Linear
pLEX-MCS
pLVX-TRE3G-mCherry
pRSI16cb-U6-sh-13kCB18-HTS6-UbiC-TagRFP-2A-Puro
pAdenoX-ZsGreen1 Linear
pLHCX
pLVX-TRE3G-ZsGreen1
pRSI16-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pB-RS
pLKO.1
pLX301
pRSI17cb-U6-sh-13kCB18-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Hygro
pLKO.1 puro
pLX302
pRSI17-U6-sh-HTS6-UbiC-TagGFP2-2A-Puro
pBABE-Neo
pLMN-ZsGreen-Neomycin
pLX303
pRSIT12-U6Tet-sh-HTS4-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Puro
pLMPd-Ametrine
pLX304
pRSIT16-U6Tet-sh-HTS6-CMV-TetRep-2A-TagRFP-2A-Puro
pBABE-Zeo
pLNCX2
pLX304-V5-Blast-Empty
pRSIT17-U6Tet-sh-HTS6-CMV-TetRep-2A-TagGFP2-2A-Puro
pCMV-VSV-G
pLP VSVG
pLXIN
pRSITEP-U6Tet-sh-noHTS-EF1-TetRep-2A-Puro
pGFP-B-RS
pLP1
pLXSN
pRSITUR-U6Tet-sh-HTS3-UbiC-TetRep-2A-TagRFP
pGFP-C-shLenti
pLP2
pMD2.G
pShuttle2
pGFP-V-RS
pLPCX
pMSCV16URP-U6-sh-HTS6-UbiC-TagRFP-2A-Puro
pSIREN-RetroQ
pGIPZ
pLVX-EF1alpha-IRES-mCherry
pMSCVhyg
pSIREN-RetroQ-DsRed-Express
pLenti-C-GFP
pLVX-EF1alpha-IRES-Puro
pMSCVneo
pSIREN-RetroQ-TetH
pLenti-C-mGFP
pLVX-EF1alpha-IRES-ZsGreen1
pMSCVpuro
pSIREN-RetroQ-TetP
pLenti-C-Myc-DDK
pLVX-EF1alpha-Tet3G
pQC-tTS-IN
pSIREN-RetroQ-ZsGreen1
pLenti-C-Myc-DDK-IRES-BSD
pLVX-Het-1
pQCXIH
psPAX2
pLenti-C-Myc-DDK-IRES-GFP
pLVX-Het-2
pQCXIN
pTRE3G-Hom1
pLenti-C-Myc-DDK-IRES-Neo
pLVX-Het-Mem1
pQCXIP
pTRIPZ
pLenti-C-Myc-DDK-IRES-Puro
pLVX-Het-Nuc1
pQCXIX
ptTS-Neo
pLenti-C-Myc-DDK-IRES-tRFP
pLVX-Hom-1
pRetro-Lib
pZIP-hCMV-RFP-Puro
pLenti-C-tRFP
pLVX-Hom-Mem1
pRetroQ-AcGFP1-C1
pZIP-hCMV-ZSGreen-Puro
pLenti-C-tYFP
pLVX-Hom-Nuc1
pRetroQ-AcGFP1-N1
pZIP-hEF1-alpha-RFP-Puro
pLenti-EF1a-C-mGFP
pLVX-IRES-Hyg
pRetroQ-DsRed-Monomer-C1
pZIP-hEF1-alpha-ZsGreen-Puro
pLenti-EF1a-C-Myc-DDK-IRES-Puro
pLVX-IRES-mCherry
pRetroQ-DsRed-Monomer-N1
pZIP-mCMV-RFP-Puro
pLenti-EF1a-C-tGFP
pLVX-IRES-Neo
pRetroQ-mCherry-C1
pZIP-mCMV-ZsGreen-Puro
pLenti-N-tGFP
pLVX-IRES-Puro
pRetroQ-mCherry-N1
pZIP-mEF1-alpha-RFP-Puro
pLenti-N-tRFP
pLVX-IRES-tdTomato
pRetroX-IRES-DsRedExpress
pZIP-mEF1-alpha-ZsGreen-Puro
pLenti-N-tYFP
pLVX-IRES-ZsGreen1
pRetroX-IRES-ZsGreen1
pZIP-SFFV-RFP-Puro
pLenti3.3 TR
pLVX-PTuner
pRetroX-PTuner
pZIP-SFFV-ZsGreen-Puro
pLenti4 BLOCK-iT-DEST
pLVX-PTuner Green
pRetroX-PTuner IRES
pZIP-TRE3G-ZsGreen-Puro
pLenti4 V5-DEST
pLVX-PTunerC
pRetroX-Tet-Off Advanced
pSIREN-Shuttle (linearized)
pLenti6.2 C-Lumio V5-DEST
pLVX-Puro
pRetroX-Tet-On Advanced
scramble shRNA

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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